Extended Data Fig. 4: Mapping of ARMC9 and TOGARAM1 interaction.
a, Schematic representation of different ARMC9 constructs. Vertical lines indicate Joubert syndrome-linked point mutations. b, Histogram plots of fluorescent intensities of single GFP molecules, GFP-EB3 dimers, and full-length GFP-ARMC dimers (left) or GFP-ARMC9 ΔH molecules immobilized in separate chambers of the same coverslips. Number of molecules analyzed, left, right: GFP, n=18415, 14914; GFP-EB3, n= 132826, 163959; GFP-ARMC9 FL, n=99873; GFP-ARMC9 ΔH, 55479. c, Co-immunoprecipitation of full-length ARMC9 with indicated proteins. Assays were repeated independently at least two times. d, Fields of view (left, scale bar 2 µm) and kymograph (right, scale bars 2 µm and 60 s) illustrating MT dynamics from GMPCPP-stabilized seed with mCherry-EB3 and indicated concentrations and colors of CTM proteins. Assays were repeated independently three times. e, f, Co-immunoprecipitation experiments of either full-length TOGARAM1 with indicated ARMC9 constructs (e) or full-length ARMC9 with indicated TOGARAM1 point mutations (f). ARMC9 and TOGARAM1 interact through ARM domain and TOG2 domain, respectively. Assays were repeated independently at least two times. g, h, Co-immunoprecipitation experiments of either full-length TOGARAM1 with indicated ARMC9 Joubert syndrome mutations (g) or full-length ARMC9 with indicated TOGARAM1 Joubert syndrome mutations (h). Assays were repeated independently at least two times.
